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| The Effects of Mytomycin-C on Stem Cells |
Elizabeth Hughes and Hannah Yewondwossen
Experiment A and Results
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Method:
Three day trial:
Using a twelve well culture dish, we placed 5ml of stem cells and medium into each well and the following amounts of Mitomycin-C were added: three wells had none; three wells had eight micrograms; three wells had two micrograms; and finally, three wells had five tenths of a microgram. These cells were then left in the incubator at a temperature of 36.8°C for a period of three days. At that time, the medium was changed to remove the Mitomycin-C. The cells were then counted. Two hundred micrograms of Trypsin 0.25% was added after rinsing the cells in 1ml of water. The cells were placed in a Carbon dioxide incubator for three minutes. The Trypsin caused the cells to contract and release themselves from the bottom and sides of the culture plate. One millilitre of medium was added to deactivate the Trypsin. The cells were then harvested through a common procedure. This consisted of using a pipette and swirling of the medium and cells. These cells were then placed in disposable tubes and the empty twelve well, cell culture plate was flushed.
Results:
Mitomycin-C has a strong effect on stem cells when given in too high a dosage. The eight microgram dosing was too large and killed all of the cells. We did not use 8mg in any further experiments.
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